High penetration prodrug compositions of mustards and mustard-related compounds

ABSTRACT

The invention provides compositions of novel high penetration compositions (HPC) or high penetration prodrugs (HPP) of mustards and mustard-related compounds, which are capable of crossing biological barriers with high penetration efficiency. The HPPs are capable of being converted to parent active drugs or drug metabolites after crossing the biological barrier and thus can render treatments for the conditions that the parent drugs or metabolites can. Additionally, the HPPs are capable of reaching areas that parent drugs may not be able to access or to render a sufficient concentration at the target areas and therefore render novel treatments. The HPPs can be administered to a subject through various administration routes, e.g., locally delivered to an action site of a condition with a high concentration or systematically administered to a biological subject and enter the general circulation with a faster rate.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a divisional application of U.S. patentapplication No. 16/209,744, filed on December 4, 2018, which is acontinuation application of U.S. patent application No. 12/418,564,filed on April 3, 2009, now U.S. Pat. No. 10,189,774, which is acontinuation-in-part application of International Application No.PCT/IB2006/053619, filed on Oct. 3, 2006 and published on Apr. 10, 2008with International Publication Number WO2008/041059, all of which areincorporated herein by reference in their entirety.

FIELD OF THE INVENTION

This invention relates to the field of pharmaceutical compositionscapable of penetrating one or more biological barriers and methods ofusing the pharmaceutical compositions for preventing, diagnosing and/ortreating condition or disease in human and animals that are treatable bymustards or mustard-related compounds. The invention also relates tomethods of using the pharmaceutical compositions for screening new drugcandidates and methods of using the pharmaceutical compositions fordiagnosing a condition in a biological subject.

BACKGROUND OF THE INVENTION

Mustards are alkylating agents and are very reactive. For example,mustards can react with DNA, RNA, and enzymes and have been used to killcancer cells in chemotherapy.

Mustards and mustard-related compounds have been used for treatment ofleukemias, breast, ovarian, and lung cancer. For example, Springer, etal. designed and synthesized many nitrogen mustard compounds for thetreatment of cancers (Springer, C. J. ea al. U.S. Pat. Nos. 6,852,755,6,916,949, and 6,005,002). Denny, et al. discussed the synthesis ofnitrobenzyl mustard quaternary salts and their use as hypoxia-selectivecytotoxic agents (Denny, W.A. et al, U.S. Pat. No. 5,691,371). Glazierdescribed prodrugs of phosphoramide mustard, isophosphoramide mustardand analogs (Glazier, A. U.S. Pat. No. 5,659,061). Farquhar designed andsynthesized novel antitumor aldophosphamide analogs (Glazier, A. U.S.Pat. No. 5,091,552). Mustards and mustard-related compounds are alsoused to treat psoriasis.

However, mustards and mustard-related compounds also cause adverse sideeffects. Common side effects of present chemotherapy using mustards ormustard-related compounds include, for example, nausea, vomiting,diarrhea, loss of appetite, hair loss, and increased susceptibility toinfection. Such side effects are often dose-dependent.

Modifications of mustards have been reported to improve their efficacyand decrease their side effects. For example, Kadow et al. described thedelivery of antitumor drugs to tumor cells by the administration of atumor-selective antibody-beta-lactamase conjugate that binds to tumorcells (Kadow, J. et al. U.S. Pat. No. 5,773,435). However, these agentsare administered orally or systematically. Oral or systematicaladministrations require a much higher plasma concentration of the activeagents to produce a therapeutically effective local concentration at theparticular site of condition or disease, e.g., cancer or infection.

Therefore, a need exists in the art for novel compositions that arecapable of being delivered efficiently and effectively to the actionsite of a condition (e.g., a disease) to prevent, reduce or treatconditions as well as minimize adverse side effects.

SUMMARY OF THE INVENTION

One aspect of the invention is directed to a high penetration prodrug(HPP) or high penetration composition (HPC) comprising a functional unitcovalently linked to a transportational unit through a linker. The terms“HPP” and “HPC” are used alone or together herein and areinterchangeable unless specifically noted.

In certain embodiments, a functional unit of a HPP or HPC comprises amoiety of an agent, wherein the efficient and effective delivery of theagent to a biological subject and/or transportation of the agent acrossone or more biological barriers are/is desired.

In certain embodiments, a functional unit may be hydrophilic,lipophilic, or amphiphilic (i.e., both hydrophilic and lipophilic). Forexample, the lipophilic nature of a function unit may be inherent orachieved by converting the hydrophilic moieties of a functional unit tolipophilic moieties.

In certain embodiments, a functional unit of a HPP or HPC comprises amoiety of a mustard or mustard-related compound. A mustard-relatedcompound is an analog of a mustard or mustard metabolite, a mustardmetabolite; or an agent that can be metabolized into a mustard ormustard metabolite after the HPP or HPC penetrates one or morebiological barriers. Examples of mustards include, but are not limitedto, nitrogen mustards, nitrobenzyl mustards, phosphoramide mustard,isophosphoramide mustards and aldophosphamide.

In certain embodiments, a transportational unit of a HPP or HPCcomprises a protonatable amine group that is capable of facilitating orenhancing the transportation or crossing of the HPP or HPC through oneor more biological barriers. In certain embodiments, the protonatableamine group is substantially protonated at the pH of the biologicalbarriers through which a HPP or HPC penetrates. In certain embodiments,the amine group can be reversibly protonated or deprotonated.

In certain embodiments, a linker covalently links the functional unit tothe transportational unit of a HPP and comprises a bond that is capableof being cleaved after the HPP penetrates across one or more biologicalbarriers. The cleavable bond comprises, for example, a covalent bond, anether, a thioether, an amide, an ester, a thioester, a carbonate, acarbamate, a phosphate or an oxime bond.

Another aspect of the invention relates to a pharmaceutical compositioncomprising at least one HPP or HPC of a mustard or mustard-relatedcompound and a pharmaceutically acceptable carrier.

Another aspect of the invention relates to a method for penetrating abiological barrier using a HPP or HPC of a mustard or mustard-relatedcompound.

Another aspect of the invention relates to a method for diagnosing theonset, development, or remission of a condition in a biological subjectby using a HPP or HPC of a mustard or mustard-related compound. Incertain embodiments, the HPP (or HPC) or the functional unit thereof isdetectable. In certain embodiments, the HPP or the functional unit ofthe HPP is inherently detectable, labeled with, or conjugated to, adetectable marker.

Another aspect of the invention relates to a method for screeningfunctional units, linkers, or transportational units for desiredcharacteristics.

Another aspect of the invention relates to a method for preventing,ameliorating, or treating a condition in a biological subject byadministering to the subject a composition in accordance with theinvention. In certain embodiments, the method relates to treating acondition in a subject treatable by mustards or mustard-relatedcompounds by administering to the subject a therapeutically effectiveamount of a HPP of a mustard or mustard-related compound, or apharmaceutical composition thereof. In certain embodiments, thepharmaceutical composition of the HPP is administrated to a biologicalsubject via various routes including, but not limted to, oral, enteral,buccal, nasal, topical, rectal, vaginal, aerosol, transmucosal,epidermal, transdermal, dermal, ophthalmic, pulmonary, subcutaneous,and/or parenteral routes. In certain preferred embodiments, thepharmaceutical composition of HPP is administered orally, transdermally,topically, subcutaneously and/or parenterally.

In accordance with the advantages of the invention, without intending tobe limited by any particular mechanism, a therapeutically effectiveamount of a HPP or HPC can be administered locally to a site ofcondition with a less dosage at a higher concentration. The advantagesof the invention also include, for example, avoidance of systematicadministration, reduction of adverse effects (e.g., pain of injection,gastrointestinal/renal effects, and other side effect), and possiblenovel treatments due to high local concentration of a HPP, HPC or activeagent. The advantages further include, for example, systematicadministration of a HPP or HPC to a biological subject to achieve fasterand more efficient bioavailability, penetration of biological barriers(e.g., the blood brain barrier) which have been difficult to cross, andnew indications as a result of passing through biological barriers.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 : Cumulative amounts of N,N-diethylaminoethyl4-[bis(2-chloroethyl)amino]benzenebutyrate.HBr (A, 20% solution),4-[bis(2-chloroethyl)am ino]-N-acetyl-L-phenylalanine N, N-diethylaminoethyl ester hydrobromide (B, 20% solution), N, N-bis(2-chloroethyl)aminophospham ide N, N-diethylam inoethyl ester hydrobrom ide (C, 10%solution), diethylam inoethyl 4-[bis(2-methylsulfonylethyl)am ino]benzenebutyrate.HCl (D, 10% solution), chlorambucil (E, 20% suspension),and melphalan (F, 20% suspension) crossing isolated human skin tissue inFranz cells (n=5). In each case, the vehicle was pH 7.4 phosphate buffer(0.2 M).

DETAILED DESCRIPTION OF THE INVENTION I. Structures of High PenetrationProdrug (HPP) or High Penetration Composition (HPC).

One aspect of the invention is directed to a high penetration prodrug(HPP) or a high penetration composition (HPC). The term “highpenetration prodrug” or “HPP” or “high penetration composition” or “HPC”as used herein refers to a composition comprising a functional unitcovalently linked to a transportational unit through a linker.

A functional unit of a HPP or HPC which comprises a moiety of a parentdrug has the properties of: 1) the delivery of the parent drug or theHPP into a biological subject and/or the transportation of the parentdrug across a biological barrier are/is desired, 2) the HPP is capableof penetrating or crossing a biological barrier, and 3) the HPP iscapable of being cleaved so as to turn the moiety of a parent drug intothe parent drug or a metabolite of the parent drug.

In certain embodiments, a functional unit may be hydrophilic,lipophilic, or amphiphilic (hydrophilic and lipophilic). The lipophilicmoiety of the function unit may be inherent or achieved by convertingits hydrophilic moieties to lipophilic moieties. For example, alipophilic moiety of a functional unit is produced by converting one ormore hydrophilic groups of the functional unit to lipophilic groups viatraditional organic synthesis. Examples of the hydrophilic groupsinclude, without limitation, carboxylic, hydroxyl, thiol, amine,phosphate/phosphonate and carbonyl groups. The lipophilic moietiesproduced via the modification of these hydrophilic groups include,without limitation, ethers, thioethers, esters, thioesters, carbonates,carbamates, amides, phosphates and oximes.

In certain embodiments, a parent drug of a HPP or HPC is selected fromthe group consisiting of a mustard and mustard-related compound. Themoiety of a mustard or mustard-related compound can be further convertedto a lipophilic moiety as described supra.

Mustards are well known in the art and are used in connection withvarious conditions. Examples of mustards and mustard-related compoundsinclude, but are not limited to, nitrogen mustards, nitrobenzylmustards, phosphoramide mustard, isophosphoramide mustards andaldophosphamide.

In one embodiment, a functional unit of a HPP of a mustard andmustard-related compound comprises a moiety having a structure selectedfrom the group consisting of Structure A and Structure B:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein:

is selected from the group consisting of Structure Y-a, Structure Y-b,

Structure Y-c, Structure Y-d, and Structure Y-e:

is selected from the group consisting of substituted and unsubstitutedaryl, Structure Ar-a, Structure Ar-b, Structure Ar-c, Structure Ar-d,Structure Ar-e, Structure Ar-f, Structure Ar-g, Structure Ar-h andStructure Ar-i:

X₁ and X₂ are independently selected from the group consisting of Cl,Br, F, I, and OSO₂R₄,

R₄ and R₆ are independently selected from the group consisting ofsubstituted and unsubstituted alkyl, substituted and unsubstitutedalkoxyl, substituted and unsubstituted perfluoroalkyl, substituted andunsubstituted alkyl halide, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl groups;

X₃-X₇ are independently selected from the group consisting of NHCOR₄,OR₄, SR₄, NHR₄, OCOR₄, R₄, substituted and unsubstituted alkoxyl,substituted and unsubstituted alkylthio, substituted and unsubstitutedalkylamino, substituted and unsubstituted alkyl halide, H, F, Cl, Br, I,NO₂, CN, CF₃, NHCOCH₃, OCH₃, SCH₃, NH₂, NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅,OC₃H₇, CH₃, C₂H₅, and C₃H₇;

m, n and z are the same or different integers;

Y₁ is selected from the group consisting of CH₂, O, S, and NH;

Y₂ and Y₃ are the same or different and each is NHCOR₄, H, OH, NHCOCH₃,NHCOC₂H₅, Cl, F, Br, or I, or taken together is =O;

Y₄ is selected from the group consisting of R₄, CH₂, —(CH₂)n-, O, S, andNH;

A is selected from the group consisting of a-amino acids, p-amino acids,and amino acids residues;

any CH₂ groups may be replaced with O, S, or NH; and when a bond is notlinked with any atom of an aryl or heteroaryl ring, the bond can be putinto any position of the ring.

In certain embodiments, the functional unit of a HPP of a mustard andmustard-related compound comprises a moiety having a structure selectedfrom the group consisting of Structure A and Structure B as definedsupra, including stereoisomers and pharmaceutically acceptable saltsthereof, wherein R₄ is selected from the group consisting of H,substituted and unsubstituted 1 to 20 carbon atoms alkyl, substitutedand unsubstituted 1 to 20 carbon atoms alkoxyl, substituted andunsubstituted 1 to 20 carbon atoms alkenyl, substituted andunsubstituted 1 to 20 carbon atoms perfluoroalkyl, substituted andunsubstituted 1 to 20 carbon atoms alkyl halide, substituted andunsubstituted 1 to 20 carbon atoms alkynyl, substituted andunsubstituted 1 to 20 carbon atoms aryl, and substituted andunsubstituted 1 to 20 carbon atoms heteroaryl moieties.

In certain embodiments, the functional unit of a HPP of a mustard andmustard-related compound comprises a moiety having a structure ofStructure B as defined supra, including stereoisomers andpharmaceutically acceptable salts thereof, wherein m is selected fromthe group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 . . .and 100.

In certain embodiments, the functional unit of a HPP of a mustard andmustard-related compound comprises a moiety having a structure ofStructure A as defined supra, including stereoisomers andpharmaceutically acceptable salts thereof, wherein n is selected fromthe group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20 . . . , and 100.

As used herein, the term “pharmaceutically acceptable salt” means thosesalts of compounds of the invention that are safe for application in asubject. Pharmaceutically acceptable salts include salts of acidic orbasic groups present in compounds of the invention. Pharmaceuticallyacceptable acid addition salts include, but are not limited to,hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate,phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate,citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate,maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate,formate, benzoate, glutamate, methanesulfonate, ethanesulfonate,benzensulfonate, p-toluenesulfonate and pamoate (i.e.,1,11-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Certain compounds ofthe invention can form pharmaceutically acceptable salts with variousamino acids. Suitable base salts include, but are not limited to,aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, anddiethanolamine salts. For a review on pharmaceutically acceptable saltssee BERGE ET AL., 66 J. PHARM. SCI. 1-19 (1 977), incorporated herein byreference.

As used herein, unless specified otherwise, the term “alkyl” means abranched or unbranched, saturated or unsaturated, monovalent ormultivalent hydrocarbon group. Examples of alkyl include, but are notlimited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl,pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, ethenyl,propenyl, butenyl, isobutenyl, pentenyl, hexenyl, heptenyl, octenyl,nonenyl, decenyl, undecenyl, dodecenyl, ethynyl, propynyl, butynyl,isobutynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl,undecynyl, dodecynyl, methylene, ethylene, propylene, isopropylene,butylene, isobutylene, t-butylene, pentylene, hexylene, heptylene,octylene, nonylene, decylene, undecylene and dodecylene, In certainembodiments, the hydrocarbon group contains 1 to 30 carbons. In certainembodiments, the hydrocarbon group contains 1 to 20 carbons

As used herein, unless specified otherwise, the term “cycloalkyl” meansan alkyl which contains at least one ring and no aromatic rings.Examples of cycloalkyl include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl. In certainembodiments, the hydrocarbon chain contains 1 to 30 carbons. In certainembodiments, the hydrocarbon group contains 1 to 20 carbons.

As used herein, unless specified otherwise, the term “heterocycloalkyl”means a cycloalkyl wherein at least one ring atom is a non-carbon atom.Examples of the non-carbon ring atom include, but are not limited to, S,O and N.

As used herein, unless specified otherwise, the term “alkoxyl” means analkyl, cycloalkyl or heterocycloalkyl, which contains one or more oxygenatoms. Examples of alkoxyl include, but are not limited to, —CH₂—OH,—OCH₃, —O-alkyl, -alkyl-OH, -alkyl-O-alkyl-, wherein the two alkyls canbe the same or different.

As used herein, unless specified otherwise, the term “alkyl halide”means an alkyl, cycloalkyl or heterocycloalkyl, which contains one ormore halogen atoms, wherein the halogen atoms can be the same ordifferent. The term “halogen” means fluorine, chlorine, bromine oriodine. Examples of alkyl halide include, but are not limited to,-alkyl-F, -alkyl-Cl, -alkyl-Br, -alkyl-I, -alkyl(F)-, -alkyl(CI)-,-alkyl(Br)- and -alkyl(I)-.

As used herein, unless specified otherwise, the term “alkylthio” meansan alkyl, cycloalkyl or heterocycloalkyl, which contains one or moresulfur atoms. Examples of alkylthio include, but are not limited to,—CH₂—SH, —SCH₃, -S-alkyl, -alkyl-SH, -alkyl-S-alkyl-, wherein the twoalkyls can be the same or different.

As used herein, unless specified otherwise, the term “alkylamino” meansan alkyl, cycloalkyl or heterocycloalkyl, which contains one or morenitrogen atoms. Examples of alkylamino include, but are not limited to,—CH₂—NH, —NCH₃, -N(alkyl)-alkyl, -N-alkyl, -alkyl-NH₂, -alkyl-N-alkyland -alkyl-N(alkyl)-alkyl wherein the alkyls can be the same ordifferent.

As used herein, unless specified otherwise, the term “perfluoroalkyl”means an alkyl, cycloalkyl or heterocycloalkyl, which contains one ormore fluoro group, including, without limitation, perfluoromethyl,perfluoroethyl, perfluoropropyl.

As used herein, unless specified otherwise, the term “aryl” means achemical structure comprising one or more aromatic rings. In certainembodiments, the ring atoms are all carbon. In certain embodiments, oneor more ring atoms are non-carbon, e.g. oxygen, nitrogen, or sulfur.Examples of aryl include, without limitation, phenyl, benzyl,naphthalenyl, anthracenyl, pyridyl, quinoyl, isoquinoyl, pyrazinyl,quinoxalinyl, acridinyl, pyrimidinyl, quinazolinyl, pyridazinyl,cinnolinyl, imidazolyl, benzimidazolyl, purinyl, indolyl, furanyl,benzofuranyl, isobenzofuranyl, pyrrolyl, indolyl, isoindolyl,thiophenyl, benzothiophenyl, pyrazolyl, indazolyl, oxazolyl,benzoxazolyl, isoxazolyl, benzisoxazolyl, thiaxolyl and benzothiazolyl.

In certain embodiments, a transportational unit of a HPP comprises aprotonatable amine group that is capable of facilitating thetransportation or crossing of the HPP through one or more biologicalbarriers (e.g., >about 50 times, >about 100 times, >about 300times, >about 500 times, >about 1,000 times faster than the parentdrug). In certain embodiments, the protonatable amine group issubstantially protonated at a physiological pH. In certain embodiments,the amine group can be reversibly protonated. In certain embodiments,the transportational unit may or may not be cleaved from the functionalunit after the penetration of HPP through one or more biologicalbarriers.

In certain embodiments, the protonatable amine group is selected fromthe group consisting of pharmaceutically acceptable substituted andunsubstituted primary amine groups, pharmaceutically acceptablesubstituted and unsubstituted secondary amine groups, andpharmaceutically acceptable substituted and unsubstituted tertiary aminegroups.

In certain embodiments, the protonatable amine group is selected fromthe group consisting of Structure Na, Structure Nb, Structure Nc,Structure Nd, Structure Ne, Structure Nf, Structure Ng, Structure Nh,Structure Ni, Structure Nj, Structure Nk, Structure NI, Structure Nm,Structure Nn, Structure No, Structure Np, Structure Nq and Structure Nr:

including stereoisomers and pharmaceutically acceptable salts thereof.

As used herein, unless specified otherwise, each R₁₁-R₁₆ isindependently selected from the group consisting of nothing, H,CH₂COOR₁₁, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl, substituted and unsubstituted alkoxyl,substituted and unsubstituted alkylthio, substituted and unsubstitutedalkylamino, substituted and unsubstituted perfluoroalkyl, andsubstituted and unsubstituted alkyl halide, wherein any carbon orhydrogen may be further independently replaced with O, S, P, NRii, orany other pharmaceutically acceptable groups.

In certain embodiments, a linker covalently linking a functional unitand a transportational unit of a HPP comprises a bond that is capable ofbeing cleaved after the HPP penetrates across one or more BBs. Thecleavable bond comprises, for example, a covalent bond, an ether,thioether, amide, ester, thioester, carbonate, carbamate, phosphate oroxime bond.

In certain embodiments, a HPP of a mustard and mustard-related compoundhas the following Structure L:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein:

F is a functional unit of a HPP of a mustard or mustard-relatedcompound. Examples of F include Structure A and Structure B as definedsupra;

T is a transportational unit of a HPP of a mustard or mustard-relatedcompound. For example, T is selected from the group consisting ofStructure Na, Structure Nb, Structure Nc, Structure Nd, Structure Ne,Structure Nf, Structure Ng, Structure Nh, Structure Ni, Structure Nj,Structure Nk, Structure NI, Structure Nm, Structure Nn, Structure No,Structure Np, Structure Nq and Structure Nr as defined supra;

Li is selected from the group consisting of nothing, O, S, -N(L₃)-,-NL₃)-CH₂—O, -NL₃)-CH₂-NL₅)-, —O—CH₂—O—, —O—CHL₃)—O—, and -S-CHL₃)-O-;L₂ is selected from the group consisting of nothing, O, S, -NL₃)-,-NL₃)-CH₂-O, -NL₃)-CH₂-NL₅)-, —O—CH₂—O—, -O-CHL₃)-O, -S-CHL₃)-O-,-0-L₃-, -N-L₃-, -S-L₃-, -NL₃)-L₅- and L₃;

L₄ is selected from the group consisting of C═O, C═S, each L₃ and L₅ isindependently selected from the group consisting of nothing, H,CH₂COOL₆, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl, substituted and unsubstituted alkoxyl,substituted and unsubstituted alkylthio, substituted and unsubstitutedalkylamino, substituted and unsubstituted perfluoroalkyl, andsubstituted and unsubstituted alkyl halide, wherein any carbon orhydrogen may be further independently replaced with 0, S, P, NL₃, or anyother pharmaceutically acceptable groups;

L₆ is selected from the group consisting of H, OH, Cl, F, Br, I,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, and substituted and unsubstituted heterocycloalkyl,substituted and unsubstituted aryl, substituted and unsubstitutedheteroaryl, substituted and unsubstituted alkoxyl, substituted andunsubstituted alkylthio, substituted and unsubstituted alkylamino,substituted and unsubstituted perfluoroalkyl, and substituted andunsubstituted alkyl halide, wherein any carbon or hydrogen may befurther independently replaced with O, S, N, P(O)OL₆, CH=CH, CEC, CHL₆,CL₆L₇, aryl, heteroaryl, or cyclic groups; and

L₇ is selected from the group consisting of H, OH, Cl, F, Br, I,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, and substituted and unsubstituted heterocycloalkyl,substituted and unsubstituted aryl, substituted and unsubstitutedheteroaryl, substituted and unsubstituted alkoxyl, substituted andunsubstituted alkylthio, substituted and unsubstituted alkylamino,substituted and unsubstituted perfluoroalkyl, and substituted andunsubstituted alkyl halide, wherein any carbon or hydrogen may befurther independently replaced with O, S, N, P(O)OL₆, CH═CH, CEC, CHL₆,CL₆L₇, aryl, heteroaryl, or cyclic groups.

In certain embodiments, a HPP or HPC of a mustard or mustard-relatedcompound comprises the structure of Structure L, including stereoisomersand pharmaceutically acceptable salts thereof, wherein F, Li, L₂ and Tare defined as supra, and L₄ is C═O.

Examples of HPPs of mustards and mustard-related compounds.

In certain embodiments, a HPP of a mustard and mustard-related compoundincludes a compound having the formula of Structure 1 or Structure 2,including stereoisomers and pharmaceutically acceptable salts thereof,wherein Structure 1 is selected from the group consisting of Structure1a, Structure 1b, Structure lc, Structure 1d, Structure 1e, Structure1f, Structure 1g, Structure 1h, Structure 1i, Structure 1j, Structure 1k, Structure 1l, Structure 1m, Structure 1n, Structure 1o, Structure 1p,Structure 1q, and Structure 1r; and Structure 2 is selected from thegroup consisting of Structure 2a, Structure 2b, Structure 2c, Structure2d, Structure 2e, Structure 2f, Structure 2g, Structure 2h, Structure2i, Structure 2j, Structure 2k, Structure 21, Structure 2m, Structure2n, Structure 2o, Structure 2p, Structure 2q, and Structure 2r:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein,

m and X₁-X₄ are defined the same as supra;

R , R₁, R₂, R₅ and R₁₁-R₁₄ are independently selected from the groupconsisting of substituted and unsubstituted 0-20 carbon atoms alkyl,substituted and unsubstituted 1-20 carbon atoms alkoxyl, substituted andunsubstituted 1-20 carbon atoms perfluoroalkyl, substituted andunsubstituted 1-20 carbon atoms alkyl halide, substituted andunsubstituted 2-20 carbon atoms alkenyl, substituted and unsubstituted2-20 carbon atoms alkynyl, substituted and unsubstituted 6-20 carbonatoms aryl, and substituted and unsubstituted 2-20 carbon atomsheteroaryl moieties which are pharmaceutically acceptable, wherein anyCH₂ may be replaced with O, S, NR₅, or other groups;

X is selected from the group consisting of O, S, NR₅, and NH; any CH₂groups may be replaced with O, S, or NH; and when a bond is not linkedwith an atom of an aryl or heteroaryl ring, the bond can be put into anyposition of the ring.

As used herein, the term “HA” is nothing or a pharmaceuticallyacceptable acid, e.g. hydrochloride, hydrobromide, hydroiodide, nitricacid , sulfic acid, bisulfic acid, phosphoric acid, phosphorous acid,phosphonic acid, isonicotinic acid, acetic acid, lactic acid, salicylicacid, citric acid, tartaric acid, pantothenic acid, bitartaric acid,ascorbic acid, succinic acid, maleic acid, gentisinic acid, fumaricacid, gluconic acid, glucaronic acid, saccharic acid, formic acid,benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid,benzensulfonic acid, p-toluenesulfonic acid and pamoic acid.

II. Pharmaceutical Compositions Comprising HPPs

Another aspect of the invention relates to a pharmaceutical compositioncomprising at least one HPP of a mustard or mustard-related compound anda pharmaceutically acceptable carrier.

The term “pharmaceutically acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting a HPP from one location,body fluid, tissue, organ (interior or exterior), or portion of thebody, to another location, body fluid, tissue, organ, or portion of thebody.

Each carrier is “pharmaceutically acceptable” in the sense of beingcompatible with the other ingredients, e.g., a HPP, of the formulationand suitable for use in contact with the tissue or organ of a biologicalsystem without excessive toxicity, irritation, allergic response,immunogenicity, or other problems or complications, commensurate with areasonable benefit/risk ratio.

Some examples of materials which can serve aspharmaceutically-acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) alcohol, such as ethyl alcohol and propane alcohol; (20)phosphate buffer solutions; and (21) other non-toxic compatiblesubstances employed in pharmaceutical formulations such as acetone.

The pharmaceutical compositions may contain pharmaceutically acceptableauxiliary substances as required to approximate physiological conditionssuch as pH adjusting and buffering agents, toxicity adjusting agents andthe like, for example, sodium acetate, sodium chloride, potassiumchloride, calcium chloride, sodium lactate and the like.

In one embodiment, the pharmaceutically acceptable carrier is an aqueouscarrier, e.g. buffered saline and the like. In certain embodiments, thepharmaceutically acceptable carrier is a polar solvent, e.g. acetone andalcohol.

The concentration of HPP in these formulations can vary widely, and willbe selected primarily based on fluid volumes, viscosities, body weightand the like in accordance with the particular mode of administrationselected and the biological system's needs. For example, theconcentration can be 0.0001% to 100%, 0.001% to 50%, 0.01% to 30%, 0.1%to 10% wt.

The compositions of the invention can be administered for prophylactic,therapeutic, and/or hygienic use. Such administration can be topical,mucosal, e.g., oral, nasal, vaginal, rectal, parenteral, transdermal,subcutaneous, intramuscular, intravenous, via inhalation, ophthalmic andother convenient routes. The pharmaceutical compositions can beadministered in a variety of unit dosage forms depending upon the methodof administration. For example, unit dosage forms suitable for oraladministration include powder, tablets, pills, capsules and lozenges.

Thus, a typical pharmaceutical composition for intravenousadministration would be about 10⁻⁹ g to about 100 g, about 10⁻⁶ g toabout 100 g, about 0.001 g to about 100 g, about 0.01 g to about 10 g,or about 0.01 g to about 1 g per subject per day. Dosages from about0.01 mg, up to about 5 g, per subject per day may be used. Actualmethods for preparing parenterally administrable compositions will beknown or apparent to those skilled in the art and are described in moredetail in such publications as Remington's Pharmaceutical Science, 15thed., Mack Publishing Company, Easton, Pa. (1980).

III. Applications of HPPs

i) Methods for penetrating a biological barrier.

Another aspect of the invention relates to a method of using acomposition of the invention in penetrating one or more biologicalbarriers in a biological subject. The method comprises a step ofadministering to a biological subject a HPP or a mustard ormustard-related compound, or a pharmaceutical composition thereof. Inone embodiment, a HPP exhibits more than about 50 times orhigher, >about 100 times or higher, >about 200 time higher, >about 300times or higher, >about 500 times or higher, >about 1,000 times orhigher penetration rate through one or more biological barriers than itsparent drug.

The term “biological barrier” as used herein refers to a biologicallayer that separates an environment into different spatial areas orcompartments, which separation is capable of modulating (e.g.restricting, limiting, enhancing or taking no action in) the passingthrough, penetrating or translocation of substance or matter from onecompartment/area to another. The different spatial areas or compartmentsas referred to herein may have the same or different chemical orbiological environment(s). The biological layer as referred hereinincludes, but is not limited to, a biological membrane, a cell layer, abiological structure, an inner surface of subjects, organisms, organs orbody cavities, an external surface of subjects, organisms, organs orbody cavities, or any combination or plurality thereof.

Examples of a biological membrane include a lipid bilayer structure,eukaryotic cell membrane, prokaryotic cell membrane, and intracellularmembrane (e.g., nucleus or organelle membrane, such as membrane orenvelope of Golgi apparatus, rough and smooth endoplasmic reticulum(ER), ribosomes, vacuoles, vesicles, liposomes, mitochondria, lysosome,nucleus, chloroplasts, plastids, peroxisomes or m icrobodies).

The lipid bilayer referred to herein is a double layer of lipid-classmolecules, including, but not limited to, phospholipids and cholesterol.In a particular embodiment, lipids for bilayer are amphiphilic moleculesconsisting of polar head groups and non-polar fatty acid tails. Thebilayer is composed of two layers of lipids arranged so that theirhydrocarbon tails face one another to form an oily core held together bythe hydrophobic effect, while their charged heads face the aqueoussolutions on either side of the membrane. In another particularembodiment, the lipid bilayer may contain one or more embedded proteinand/or sugar molecule(s).

Examples of a cell layer include a lining of eukaryotic cells (e.g.,epithelium, lamina propria and smooth muscle or muscularis mucosa (ingastrointestinal tract)), a lining of prokaryotic cells (e.g., surfacelayer or S-layer which refers to a two dimensional structuremonomolecular layer composed of identical proteins or glycoproteins,specifically, an S-layer refers to a part of a cell envelope commonlyfound in bacteria and archaea), a biofilm (a structured community ofmicroorganisms encapsulated within a self-developed polymeric matrix andadherent to a living or inert surface), and a plant cell layer (e.g.,empidermis). The cells may be normal cells or pathological cells (e.g.disease cells, cancer cells).

Examples of biological structures include structures sealed by tight oroccluding junctions which provide a barrier to the entry of toxins,bacteria and viruses, e.g. the blood milk barrier and the blood brainbarrier (BBB). In particular, BBB is composed of an impermeable class ofendothelium, which presents both a physical barrier through tightjunctions adjoining neighboring endothelial cells and a transportbarrier comprised of efflux transporters. The biological structure mayalso include a mixture of cells, proteins and sugars (e.g. blood clots).

Examples of the inner surface of subjects, organisms, organs or bodycavities include buccal mucosa, esophageal mucosa, gastric mucosa,intestinal mucosa, olfactory mucosa, oral mucosa, bronchial mucosa,uterine mucosa and endometrium (the mucosa of the uterus, inner layer ofthe wall of a pollen grain or the inner wall layer of a spore), or acombination or plurality thereof.

Examples of the external surface of subjects, organisms, organs or bodycavities include capillaries (e.g. capillaries in the heart tissue),mucous membranes that are continuous with skin (e.g. such as at thenostrils, the lips, the ears, the genital area, and the anus), outersurface of an organ (e.g. liver, lung, stomach, brain, kidney, heart,ear, eye, nose, mouth, tongue, colon, pancreas, gallbladder, duodenum,rectum stomach, colonrectum, intestine, vein, respiratory system,vascular, the anorectum and pruritus ani), skin, cuticle (e.g., deadlayers of epidermal cells or keratinocytes or superficial layer ofoverlapping cells covering the hair shaft of an animal, a multi-layeredstructure outside the epidermis of many invertebrates, plant cuticles orpolymers cutin and/or cutan), external layer of the wall of a pollengrain or the external wall layer of a spore), and a combination orplurality thereof.

In addition, a biological barrier further includes a sugar layer, aprotein layer or any other biological layer, or a combination orplurality thereof. For example, skin is a biological barrier that has aplurality of biological layers. A skin comprises an epidermis layer(outer surface), a demis layer and a subcutaneous layer. The epidermislayer contains several layers including a basal cell layer, a spinouscell layer, a granular cell layer, and a stratum corneum. The cells inthe epidermis are called keratinocytes. The stratum corneum (“hornylayer”) is the outmost layer of the epidermis, wherein cells here areflat and scale-like (“squamous”) in shape. These cells contain a lot ofkeratin and are arranged in overlapping layers that impart a tough andoilproof and waterproof character to the skin's surface.

ii) Methods for diagnosing a condition in a biological system.

Another aspect of the invention relates to a method of using acomposition of the invention in diagnosing a condition in a biologicalsystem. The method comprises the following steps:

-   1) administrating a composition comprising a HPP of a mustard or    mustard-related compound to the biological subject;-   2) detecting the presence, location or amount of the HPP, the    functional unit of the HPP or a metabolite thereof in the biological    subject; and-   3) determining a condition in the biological system.

In certain embodiments, the HPP (or the agent cleaved from the HPP)aggregates in the site of action where a condition occurs. In certainembodiments, the presence, location or amount of the functional unit ofthe HPP is also detected. In certain embodiments, the onset,development, progress, or remission of a condition (e.g., cancer)associated is also determined.

In certain embodiments, the HPP is labeled with or conjugated to adetectable agent. Alternatively, the HPP is prepared to includeradioisotopes for detection. Numerous detectable agents are availablewhich can be generally grouped into the following categories:

(a) Radioisotopes, such as ³⁵S, ¹⁴C, ¹³C, ¹⁵N, ¹²⁵I, ³H, and ¹³¹I. Thediagnostic agent can be labeled with the radioisotope using thetechniques known in the art and radioactivity can be measured usingscintillation counting; in addition, the diagnostic agent can be spinlabeled for electron paramagnetic resonance for carbon and nitrogenlabeling.

(b) Fluorescent agents such as BODIPY, BODIPY analogs, rare earthchelates (europium chelates), fluorescein and its derivatives, FITC, 5,6carboxyfluorescein, rhodamine and its derivatives, dansyl, Lissamine,phycoerythrin, green fluorescent protein, yellow fluorescent protein,red fluorescent protein and Texas Red. Fluorescence can be quantifiedusing a fluorometer.

(c) Various enzyme-substrate agents, such luciferases (e.g., fireflyluciferase and bacterial luciferase), luciferin,2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidasesuch as horseradish peroxidase (HRPO), alkaline phosphatase,13-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g.,glucose oxidase, galactose oxidase, and glucose-6-phosphatedehydrogenase), heterocyclic oxidases (such as uricase and xanthineoxidase), lactoperoxidase, microperoxidase, and the like. Examples ofenzyme-substrate combinations include, for example: (i) Horseradishperoxidase (HRPO) with hydrogen peroxidase as a substrate, wherein thehydrogen peroxidase oxidizes a dye precursor (e.g., orthophenylenediamine (OPD) or 3,3′,5,5′-tetramethyl benzidine hydrochloride (TMB));(ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate aschromogenic substrate; and (iii) β-D-galactosidase (β-D-Gal) with achromogenic substrate (e.g., p-nitrophenyl-β-D-galactosidase) orfluorogenic substrate 4-methylumbelliferyl-β-D-galactosidase.

In certain embodiments, the detectable agent is not necessarilyconjugated to the diagnostic agent but is capable of recognizing thepresence of the diagnostic agent and the diagnostic agent can bedetected.

In certain embodiments, the HPP of the invention can be provided in akit, i.e., a packaged combination of reagents in predetermined amountswith instructions for performing the diagnostic assay. Where the HPP islabeled with an enzyme, the kit will include substrates and cofactorsrequired by the enzyme (e.g., a substrate precursor which provides thedetectable chromophore or fluorophore). In addition, other additives maybe included such as stabilizers, buffers (e.g., a block buffer or lysisbuffer) and the like. The relative amounts of the various reagents maybe varied widely to provide for concentrations in solution of thereagents which substantially optimize the sensitivity of the assay.Particularly, the reagents may be provided as dry powders, usuallylyophilized, including excipients which on dissolution will provide areagent solution having the appropriate concentration.

iii) Methods for Screening a Substance for a Desired Character

Another aspect of the invention relates to a method of screening a HPPfor a desired character.

In certain embodiments, the method comprises:

-   1) covalently linking a test functional unit to a transportational    unit through a linker to form a test composition (or covalently    linking a functional unit to a test transportational unit through a    linker, or covalently linking a functional unit to a    transportational unit through a test linker)-   2) administrating the test composition to a biological system; and-   3) determining whether the test composition has the desired nature    or character.

In one embodiment, a desired character may include, for example, 1) theability of a test functional unit to form a high penetration compositionor convert back to a parent drug, 2) the penetration ability and/or rateof a test composition, 3) the efficiency and/or efficacy of a testcomposition, 4) the transportational ability of a test transportationalunit, and 5) the cleavability of a test linker.

iv) Methods for Treating a Condition in a Biological Subject

Another aspect of the invention relates to a method of using acomposition of the invention in treating a condition in a biologicalsystem. The method comprises administrating the pharmaceuticalcomposition to the biological system.

The term “treating” as used herein means curing, alleviating,inhibiting, or preventing. The term “treat” as used herein means cure,alleviate, inhibit, or prevent. The term “treatment” as used hereinmeans cure, alleviation, inhibition or prevention.

The term “biological system,” “biological subject” or “subject” as usedherein means an organ, a group of organs that work together to perform acertain task, an organism, or a group of organisms. The term “organism”as used herein means an assembly of molecules that function as a more orless stable whole and has the properties of life, such as animal, plant,fungus, or micro-organism.

The term “animal” as used herein means an eukaryotic organismcharacterized by voluntary movement. Examples of animal include, withoutlimitation, vertebrata (e.g. human, mammals, birds, reptiles,amphibians, fishes, marsipobranchiata and leptocardia), tunicata (e.g.thaliacea, appendicularia, sorberacea and ascidioidea), articulata (e.g.insecta, myriapoda, malacapoda, arachnida, pycnogonida, merostomata,crustacea and annelida), gehyrea (anarthropoda), and helminthes (e.g.rotifera).

The term “plant” as used herein means organisms belonging to the kindomPlantae. Examples of plant include, without limitation, seed plants,bryophytes, ferns and fern allies. Examples of seed plants include,without limitation, cycads, ginkgo, conifers, gnetophytes, angiosperms.Examples of bryophytes include, without limitation, liverworts,hornworts and mosses. Examples of ferns include, without limitation,ophioglossales (e.g. adders-tongues, moonworts, and grape-ferns),marattiaceae and leptosporangiate ferns. Examples of fern alliesinclude, without limitation, lycopsida (e.g. clubmosses, spikemosses andquillworts), psilotaceae (e.g. lycopodiophyta and whisk ferns) andequisetaceae (e.g. horsetails).

The term “fungus” as used herein means a eukaryotic organism that is amember of the kingdom Fungi. Examples of fungus include, withoutlimitation, chytrids, blastocladiomycota, neocallimastigomycota,zygomycota, glomeromycota, ascomycota and basidiomycota.

The term “micro-organism” as used herein means an organism that ismicroscopic (e.g. with length scale of micrometer), Examples ofmicro-organism include, without limitation, bacteria, fungi, archaea,protists and microscopic plants (e.g. green algae) and microscopicanimals (e.g. plankton, planarian and amoeba).

Some examples of the conditions the method can treat include conditionsthat can be treated by the parent drug of the HPP.

v). Methods of using HPPs of Mustards and Mustard-Related Compounds andPharmaceutical Compositions thereof in Treatments.

Another aspect of the invention relates to a method of using HPPs ofmustards or mustard-related compounds, or pharmaceutical compositionsthereof in treating a condition in a biological system or subject byadministrating a HPP of a mustard or mustard-related compound, or apharmaceutical compositions thereof to the biological system or subject.

Examples of the conditions that can be treated by the method includepsoriasis and tumor, e.g., benign tumor, breast cancer, colon-rectumcancer, oral cancer, lung or other respiratory system cancers, skincancers, uterus cancer, pancreatic cancer, prostate cancer, genitalcancer, urinary organs cancers, leukemia or other blood and lymphtissues cancer.

In certain embodiments, a method of treating a condition in a subjectamelioratable or treatable with mustards or mustard-related compoundscomprises administering a therapeutic effective amount of a HPP of amustard or mustard-related compound, or a pharmaceutical compositionthereof to the subject.

A HPP or a pharmaceutical composition thereof can be administered to abiological system by any administration route known in the art,including without limitation, oral, enteral, buccal, nasal, topical,rectal, vaginal, aerosol, transmucosal, epidermal, transdermal, dermal,ophthalmic, pulmonary, subcutaneous, and/or parenteral administration.The pharmaceutical compositions can be administered in a variety of unitdosage forms depending upon the method of administration.

A parenteral administration refers to an administration route thattypically relates to injection which includes but is not limited tointravenous, intramuscular, intraarterial, intrathecal, intracapsular,intraorbital, intra cardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal, and/or intrasternal injection and/orinfusion.

A HPP or a pharmaceutical composition thereof can be given to a subjectin the form of formulations or preparations suitable for eachadministration route. The formulations useful in the methods of theinvention include one or more HPPs, one or more pharmaceuticallyacceptable carriers therefor, and optionally other therapeuticingredients. The formulations may conveniently be presented in unitdosage form and may be prepared by any methods well known in the art ofpharmacy. The amount of active ingredient which can be combined with acarrier material to produce a single dosage form will vary dependingupon the subject being treated and the particular mode ofadministration. The amount of a HPP which can be combined with a carriermaterial to produce a pharmaceutically effective dose will generally bethat amount of a HPP which produces a therapeutic effect. Generally, outof one hundred percent, this amount will range from about 1 percent toabout ninety-nine percent of the HPP, preferably from about 20 percentto about 70 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association a HPP with one or more pharmaceuticallyacceptable carriers and, optionally, one or more accessory ingredients.In general, the formulations are prepared by uniformly and intimatelybringing into association a HPP with liquid carriers, or finely dividedsolid carriers, or both, and then, if necessary, shaping the product.

Formulations suitable for oral administration may be in the form ofcapsules, cachets, pills, tablets, lozenges (using a flavored basis,usually sucrose and acacia or tragacanth), powders, granules, or as asolution or a suspension in an aqueous or non-aqueous liquid, or as anoil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup,or as pastilles (using an inert base, such as gelatin and glycerin, orsucrose and acacia) and/or as mouth washes and the like, each containinga predetermined amount of a HPP as an active ingredient. A compound mayalso be administered as a bolus, electuary, or paste.

In solid dosage forms for oral administration (e. g., capsules, tablets,pills, dragees, powders, granules and the like), the HPP is mixed withone or more pharmaceutically-acceptable carriers, such as sodium citrateor dicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,certain silicates, and sodium carbonate, (5) solution retarding agents,such as paraffin, (6) absorption accelerators, such as quaternaryammonium compounds; (7) wetting agents, such as, for example, acetylalcohol and glycerol monostearate; (8) absorbents, such as kaolin andbentonite clay; (9) lubricants, such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and (10) coloring agents. In the case of capsules,tablets and pills, the pharmaceutical compositions may also comprisebuffering agents. Solid compositions of a similar type may also beemployed as fillers in soft and hard-filled gelatin capsules using suchexcipients as lactose or milk sugars, as well as high molecular weightpolyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered peptide orpeptidomimetic moistened with an inert liquid diluent. Tablets, andother solid dosage forms, such as dragees, capsules, pills and granules,may optionally be scored or prepared with coatings and shells, such asenteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of a HPP therein using, for example,hydroxypropylmethyl cellulose in varying proportions to provide thedesired release profile, other polymer matrices, liposomes and/ormicrospheres. They may be sterilized by, for example, filtration througha bacteria-retaining filter, or by incorporating sterilizing agents inthe form of sterile solid compositions which can be dissolved in sterilewater, or some other sterile injectable medium immediately before use.These compositions may also optionally contain pacifying agents and maybe of a composition that they release the HPP(s) only, orpreferentially, in a certain portion of the gastrointestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes. The HPP canalso be in micro-encapsulated form, if appropriate, with one or more ofthe above-described excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the HPP, the liquid dosage forms may containinert diluents commonly used in the art, such as, for example, water orother solvents, solubilizing agents and emulsifiers, such as ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils(in particular, cottonseed, groundnut, corn, germ, olive, castor andsesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycolsand fatty acid esters of sorbitan, and mixtures thereof. Besides inertdiluents, the oral compositions can also include adjuvants such aswetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the HPP, may contain suspending agents as,for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitoland sorbitan esters, microcrystalline cellulose, aluminum metahydroxide,bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as asuppository, which may be prepared by mixing one or more HPPs with oneor more suitable nonirritating excipients or carriers comprising, forexample, cocoa butter, polyethylene glycol, a suppository wax or asalicylate, and which is solid at room temperature, but liquid at bodytemperature and, therefore, will melt in the rectum or vaginal cavityand release the active agent. Formulations which are suitable forvaginal administration also include pessaries, tampons, creams, gels,pastes, foams or spray formulations containing such carriers as areknown in the art to be appropriate.

Formulations for the topical or transdermal or epidermal or dermaladministration of a HPP composition include powders, sprays, ointments,pastes, creams, lotions, gels, solutions, patches and inhalants. Theactive component may be mixed under sterile conditions with apharmaceutically acceptable carrier, and with any preservatives,buffers, or propellants which may be required. The ointments, pastes,creams and gels may contain, in addition to the HPP composition,excipients, such as animal and vegetable fats, oils, waxes, paraffins,starch, tragacanth, cellulose derivatives, polyethylene glycols,silicones, bentonites, silicic acid, talc and zinc oxide, or mixturesthereof. Powders and sprays can contain, in addition to the HPPcomposition, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

A HPP or a pharmaceutical composition thereof can be alternativelyadministered by aerosol. This can be accomplished by preparing anaqueous aerosol, liposomal preparation or solid particles containing theHPPs. A nonaqueous (e. g., fluorocarbon propellant) suspension could beused. Sonic nebulizers can also be used. An aqueous aerosol is made byformulating an aqueous solution or suspension of the agent together withconventional pharmaceutically acceptable carriers and stabilizers. Thecarriers and stabilizers vary with the requirements of the particularcompound, but typically include nonionic surfactants (Tweens, Pluronics,or polyethylene glycol), innocuous proteins like serum albumin, sorbitanesters, oleic acid, lecithin, amino acids such as glycine, buffers,salts, sugars or sugar alcohols. Aerosols generally are prepared fromisotonic solutions.

Transdermal patches can also be used to deliver HPP compositions to antumor site. Such formulations can be made by dissolving or dispersingthe agent in the proper medium. Absorption enhancers can also be used toincrease the flux of the peptidomimetic across the skin. The rate ofsuch flux can be controlled by either providing a rate controllingmembrane or dispersing the peptidomimetic in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

Formulations suitable for parenteral administration comprise a HPP incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacterostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the formulations suitable for parenteral administrationinclude water, ethanol, polyols (e. g., such as glycerol, propyleneglycol, polyethylene glycol, and the like), and suitable mixturesthereof, vegetable oils, such as olive oil, and injectable organicesters, such as ethyl oleate. Proper fluidity can be maintained, forexample, by the use of coating materials, such as lecithin, by themaintenance of the required particle size in the case of dispersions,and by the use of surfactants.

Formulations suitable for parenteral administration may also containadjuvants such as preservatives, wetting agents, emulsifying agents anddispersing agents. Prevention of the action of microorganisms may beensured by the inclusion of various antibacterial and antifungal agents,for example, paraben, chlorobutanol, phenol sorbic acid, and the like.It may also be desirable to include isotonic agents, such as sugars,sodium chloride, and the like into the compositions. In addition,prolonged absorption of the injectable pharmaceutical form may bebrought about by the inclusion of agents which delay absorption such asaluminum monostearate and gelatin.

Injectable depot forms are made by forming microencapsule matrices of aHPP or in biodegradable polymers such as polylactide-polyglycolide.Depending on the ratio of the HPP to polymer, and the nature of theparticular polymer employed, the rate of drug release can be controlled.Examples of other biodegradable polymers include poly (orthoesters) andpoly (anhydrides). Depot injectable formulations are also prepared byentrapping the HPP in liposomes or microemulsions which are compatiblewith body tissue.

In certain embodiments, a HPP of a mustard or mustard-related compound,or a pharmaceutical composition thereof is delivered to a disease ortumor site in a therapeutically effective dose. As is known in the artof pharmacology, the precise amount of the pharmaceutically effectivedose of a HPP that will yield the most effective results in terms ofefficacy of treatment in a given patient will depend upon, for example,the activity, the particular nature, pharmacokinetics, pharmacodynamics,and bioavailability of a particular HPP, physiological condition of thesubject (including race, age, sex, weight, diet, disease type and stage,general physical condition, responsiveness to a given dosage and type ofmedication), the nature of pharmaceutically acceptable carriers in aformulation, the route and frequency of administration being used, andthe severity or propensity of a disease caused by pathogenic targetmicrobial organisms, to name a few. However, the above guidelines can beused as the basis for fine-tuning the treatment, e. g., determining theoptimum dose of administration, which will require no more than routineexperimentation consisting of monitoring the subject and adjusting thedosage. Remington: The Science and Practice of Pharmacy (Gennaro ed.20.sup.th edition, Williams & Wilkins Pa., USA) (2000).

IV. Advantages

In certain embodiments, since a HPP of the invention is capable ofcrossing one or more biological barriers, the HPP can be administeredlocally (e.g., topically or transdermally) to reach a location where acondition occurs without the necessity of a systematic administration(e.g., oral or parenteral administration). A local administration andpenetration of a HPP allows the HPP to reach the same level of localconcentration of an agent or drug with much less amount or dosage of HPPin comparison to a systematic administration of a parent agent or drug;alternatively, a higher level of local concentration which may not beafforded in the systematic administration, or if possible, requiressignificantly higher dosage of an agent in the systematicadministration. The high local concentration of the HPP or its parentagent if being cleaved enables the treatment of a condition moreeffectively or much faster than a systematically delivered parent agentand the treatment of new conditions that may not be possible or observedbefore. The local administration of the HPP may allow a biologicalsubject to reduce potential sufferings from a systemic administration,e.g., adverse reactions associated with the systematic exposure to theagent, gastrointestinal/renal effects. Additionally, the localadministration may allow the HPP to cross a plurality of biologicalbarriers and reach systematically through, for example, generalcirculation and thus avoid the needs for systematic administration(e.g., injection) and obviate the pain associated with the parenteralinjection.

In certain embodiments, a HPP or a pharmaceutical composition accordingto the invention can be administered systematically (e.g., orally orparenterally). The HPP or the active agent (e.g., drug or metabolite) ofthe HPP may enter the general circulation with a faster rate than theparent agent and gain faster access to the action site a condition.Additionally, the HPP can cross a biological barrier (e.g., blood brainbarrier) which has not been penetrated if a parent agent is administeredalone and thus offer novel treatment of conditions that may not bepossible or observed before.

For example, HPPs of mustards or mustard-related compounds in theinvention demonstrated high penetration rate through a biologicalbarrier (e.g., >about 10 times, >about 50 times, >about 100times, >about 200 times, >about 300 times higher than if the mustards ormustard-related compounds are administered alone). No or few adverseside effect was observed from the subjects that took mustards HPP, whileside effects (such as nausea, hair loss, and increased susceptibility toinfection) were observed from the subjects that took the parent mustardsat the similar dosage.

V. Examples

The following examples are provided to better illustrate the claimedinvention and are not to be interpreted in any way as limiting the scopeof the invention. All specific compositions, materials, and methodsdescribed below, in whole or in part, fall within the scope of theinvention. These specific compositions, materials, and methods are notintended to limit the invention, but merely to illustrate specificembodiments falling within the scope of the invention. One skilled inthe art may develop equivalent compositions, materials, and methodswithout the exercise of inventive capacity and without departing fromthe scope of the invention. It will be understood that many variationscan be made in the procedures herein described while still remainingwithin the bounds of the invention. It is the intention of the inventorsthat such variations are included within the scope of the invention.

Example 1 Preparation of a HPP from a Parent Drug

Preparation of a HPP from a Parent Drug which Contains at least oneCarboxylic Group.

In certain embodiments, a parent compound having the following StructureC:

is converted to a HPP having Structure L:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein: F, Li, L₂, L₄ and T are defined the same as supra;

In certain embodiments of the invention, a HPP having Structure L isprepared according to the conventional organic synthesis by reacting theparent compounds or derivatives of the parent compounds having StructureD (e.g. acid halides, mixed anhydrides of the parent compounds, etc.):

with compounds of Structure E (Scheme 1):

T-L₂-H

Structure E

wherein W is selected from the group consisting of OH, halogen,alkoxycarbonyl and substituted aryloxycarbonyloxy; and

F, T, Li, L₂, and L₄ are defined the same as supra.

In certain embodiments, a HPP having Structure L is prepared followingScheme 1 as described supra, wherein L₄ is C═O.

Preparation of N,N-diethylaminoethyl 4-[bis(2-chloroethyl) amino]benzenebutyrate.HBr

32.6 g (0.1 mol) of sodium 4-[bis(2-chloroethyl)amino]benzenebutyratewas dissolved in 100 ml of acetonitrile. 26 g (0.10 mol) of2-Bromo-N,N-diethylethylamine.HBr in ethyl acetate was added into thereaction mixture. The mixture was stirred for 3 h at RT. Solid isremoved by filtration. The solvents were evaporated off. The solidproduct was collected by filtration and washed with ether. After drying,it yielded 35 g of the desired product (72.3%). Hygroscopic product;Solubility in water: 300 mg/ml; Elementary analysis: C₂₀H₃₃BrCl₂N₂O₂;MW: 484.30. Calculated % C: 49.60, H: 6.87, Br: 16.50, N: 5.78, 0: 6.61; CI: 14.64; Found % C: 49.52, H: 6.89, Br: 16.55 N: 5.75, 0: 6.65; Cl:14.64. ¹H-NMR (400 MHz, D₂O): δ: 1.55 (t, 6H), 2.02(m, 2H), 2.27 (m,2H), 2.54 (m, 2H), 3.23 (m, 4H), 3.51 (m, 2H), 3.60-3.65 (m, 8H), 4.51(m, 2H), 6.55 (m, 2H), 6.95 (m, 2H).

Preparation of N,N-diethylaminoethyl 4-[bis(2-chloroethyl) amino]benzenebutyrate.HCl

(A) 30.4 g (0.1 mol) of 4-[bis(2-chloroethyl)amino]benzenebutanoic acidwas dissolved in 300 ml of chloroform. 20.6 g of N,N′-Dicyclohexylcarbodiimide was added into the reaction mixture. 11.7 gof N,N-diethylaminoethanol and 0.2 g of 4-dimethylaminopyridine wereadded into the reaction mixture. The mixture was stirred overnight at 0°C. The solid was removed by filtration. The chloroform solution waswashed with water (1×100 ml), 5% NaHCO₃ (1×100 ml) and water (3×100 ml).The organic solution was dried over anhydrous sodium sulfate. Sodiumsulfate was removed by filtration. 4 g of HCl gas in methanol (10 ml)was added into the reaction mixture with stirring. Hexane (200 ml) wasadded. The solid product was collected by filtration. After drying, ityielded 35 g of the desired product (79.6%). Hygroscopic product;Solubility in water: 300 mg/ml; Elementary analysis: C₂₀H₃₃Cl₃N₂O₂; MW:439.85. Calculated % C: 54.61, H: 7.56, N: 6.37; O: 7.27; Cl: 24.18;Found % C: 54.55; H: 7.58; N: 6.34, O: 7.29; Cl: 24.24. 1H-NMR (400 MHz,D2O): δ: 1.56 (t, 6H), 2.01(m, 2H), 2.25 (m, 2H), 2.55 (m, 2H), 3.22 (m,4H), 3.52 (m, 2H), 3.60-3.65 (m, 8H), 4.50 (m, 2H), 6.55 (m, 2H), 6.95(m, 2H).

(B) 60.8 g (0.1 mol) of 4-[bis(2-chloroethyl)amino]benzenebutanoic acidwas dissolved in 300 ml of chloroform. 20.6 g of N,N′-Dicyclohexylcarbodiimide was added into the reaction mixture. 11.7 gof N,N-diethylaminoethanol was added into the reaction mixture. Themixture was stirred overnight at 0° C. The solid was removed byfiltration. The chloroform solution was washed with water (1×100 ml), 5%NaHCO₃ (1×100 ml) and water (3×100 ml). The organic solution was driedover anhydrous sodium sulfate. Sodium sulfate was removed by filtration.4 g of HCl gas in methanol (10 ml) was added into the reaction mixturewith stirring. Hexane (200 ml) was added. The solid product wascollected by filtration. After drying, it yielded 32 g of the desiredproduct (73%). Hygroscopic product; Elementary analysis: C₂₀H₃₃Cl₃N₂O₂;MW: 439.85. Calculated % C: 54.61, H: 7.56, N: 6.37; O: 7.27; Cl: 24.18;Found % C: 54.57; H: 7.57; N: 6.34, O: 7.29; Cl: 24.23.

Preparation of 4-[bis(2-chloroethyl)amino]-N-acetyl-L-phenylalanineN,N-diethylaminoethyl ester hydrobromide

36.9 g (0.1 mol) of sodium4-[bis(2-chloroethyl)amino]-N-acetyl-L-phenylalanine was dissolved in100 ml of acetonitrile. 26 g (0.10 mol) of2-Bromo-N,N-diethylethylamine.HBr in ethyl acetate was added into thereaction mixture. The mixture was stirred for 3 h at RT. Solid isremoved by filtration. The solvents were evaporated off. The solidproduct was collected by filtration and washed with ether. After drying,it yielded 38 g of the desired product (72.1%). Hygroscopic product;Solubility in water: 300 mg/ml; Elementary analysis: C₂₁H₃₄BrCl₂N₂O₂;MW: 527.32. Calculated % C: 47.83, H: 6.50, Br: 15.15, N: 7.97, O: 9.10,CI: 13.45; Found % C: 47.77, H: 6.52, Br: 15.12 N: 7.96, O: 9.15; Cl:13.48. ¹H-NMR (400 MHz, D20): δ: 1.54 (t, 6H), 2.02(s, 3H), 3.16 (m,2H), 3.23 (m, 4H), 3.51 (m, 2H), 3.60-3.65 (m, 8H), 4.51 (m, 2H), 4.81(m, 1H), 6.55 (m, 2H), 6.95 (m, 2H).

EXAMPLE 2 HPPs of Mustards and Mustard-Related Compounds have Higher invitro Penetration Rates across Human Skin Comparing to their ParentDrugs

The penetration rates of HPPs and their parent drugs through human skinwere measured in vitro by modified Franz cells. The Franz cells had twochambers, the top sample chamber and the bottom receiving chamber. Thehuman skin tissue (360-400 pm thick) that separated the top and thereceiving chambers was isolated from the anterior or posterior thighareas.

The compound tested (2 mL, 20% in 0.2 M phosphate buffer, pH. 7.4) wereadded to the sample chamber of a Franz cell. The receiving chambercontains 10 ml of 2% bovine serum albumin in saline which was stirred at600 rpm. The amount of the tested compound penetrating the skin wasdetermined by high-performance liquid chromatography (HPLC) method. Theresults were shown in FIG. 1 . The apparent flux values of the testedcompounds were calculated from the slopes in FIG. 1 and summarized inTable 1.

Because the lowest detectable apparent flux values in this method was 1μg/cm²/h, parent drugs that showed a apparent flux value less than 1μg/cm²/h were considered as not detectable for penetrating across theskin tissue. The HPPs of these parent drugs (e.g. nitrogen mustards,nitrobenzyl mustards, phosphoramide mustard, isophosphoramide mustardsand aldophosphamide) had detectable penetration across the skin tissue.For the parent drugs that had detectable apparent flux value, their HPPshad higher apparent flux value.

TABLE 1 In vitro Penetration Rate of HPPs and their Parent Compounds mg/mg/ HPPs cm²/h Parent compounds cm²/h N,N-diethylaminoethyl 1.01chlorambucil 0.01 4-[bis(2-chloroethyl)amino] benzenebutyrate.HBr4-[bis(2-chloroethyl)amino]- 1.10 melphalan 0.01N-acetyl-L-phenylalanine N,N-diethylaminoethyl ester hydrobromideN,N-bis(2-chloroethyl) 0.85 N,N-bis(2- 0.01 aminophosphamidechloroethyl) [Dr. Yu, N,N-diethylaminoethyl ester aminophosphamideplease hydrobromide confirm.] diethylaminoethyl 4-[bis(2- 0.94 4-[bis(2-0.01 methylsulfonylethyl)amino] methylsulfonylethyl) [Dr. Yu,benzenebutyrate.HBr amino] please benzenebutanoic confirm.] acid

EXAMPLE 3 In vivo Transportation of Prodrug and Application of HPPs ofMustards and Mustard-Related Compounds in Treating Cancer

1) Blocking Human Gastric Cancer HGC-27 cell Proliferation withChlorambucil and N, N-diethylam inoethyl4-[bis(2-chloroethyl)amino]benzenebutyrate The inhibition of cellularproliferation was measured by the modified dimethyl thiazolyl diphenyltetrazolium salt (MTT)[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay,based on the ability of live cells to converting thiazolyl blue to darkblue formazan. Approximately 3500 cells of HGC-27 (in 100 μl culturesolution) were seeded into 96-well culture plates and were cultured for16 hours at 37° C. Different concentration solution (100 μl) of Taxol(positive control), chlorambucil, or N,N-diethylaminoethyl4-[bis(2-chloroethyl) amino]benzenebutyrate were added and incubationcontinued for 72 hours at 37° C. Then MTT were added and incubationcontinued at 37° C. for 4 h, and 100 μl DMSO was pipetted to solubilizethe formazan product for 30 min at room temperature. The absorbency at570 nm was measured using Bio-Rad micro-plate reader stored at −20° C.until use for electrophoresis. EC50 were calculated with the softwarePrism Graphpad.

Results: HGC-27 cell growth inhibition rates for chlorambucil at thefinal concentrations of 500, 200, 100, 75, 50, 25, 5, 2, 0.5 pm were81.0%, 47.7%, 39.2%, 34.2%, 34.9%, 25.0%, 1.6%, 0.3% and EC50 is >100μm. HGC-27 cell growth inhibition rates for N,N-diethylaminoethyl4-[bis(2-chloroethyl)amino]benzenebutyrate at the final concentrationsof 500, 200, 100, 75, 50, 25, 5, 2, 0.5 μm were 94.8% , 94.8% , 94.7%,93.4%, 81.6%, 54.6%, 46.9%, 34.7%, 25.5%, EC50 was <6μM. HGC-27 cellgrowth inhibition rates for taxol at the final concentrations of 5000nm, 500 nm, 100 nm, 50 nm, 25 nm, 10 nm, 1 nm, 0.5 nm were 95.8%, 94.7%,92.2%, 85.2%, 76.9%, 45.0%, 21.6%, 10.3% and EC50 was ˜13 nm.

2) For evaluation of antitumor activity, a human myeloma cell linederived from the ascites of a patient with multiple myeloma wasimplanted into mice.

The experiment was carried out on 11 groups of mice. Control group (A,orally), melphalan (B₁ and B₂, orally), chlorambucil (C₁ and C₂,orally), N,N-diethylaminoethyl 4-[bis(2-chloroethyl)amino]benzenebutyrate.HBr (D₁ and D₂, transdermally),4-[bis(2-chloroethyl)amino]-N-acetyl-L-phenylalanine N,N-diethylaminoethyl ester hydrobromide (Ei and E2, transdermally), anddiethylaminoethyl 4-[bis(2-methylsulfonylethyl)amino]benzenebutyrate.HCl(F₁ and F₂, transdermally). The body weight loss of mice was determinedon day 21. The results were shown in Table 2.

TABLE 2 Extension of survival period and weight loss of cancer mice byuse of mustards and their novel prodrugs. Dose Survival Life None Weight(mg/kg) Period Elongation Disease Loss Compounds perday n (days) Rate(%)Rate (%) Control (A) — 7  45.5 ± 1.6 100 0/7 1% B 1.5 mg 7  55.7 ± 1.3122 0/7 10%  B 3 mg 7  88.5 ± 1.8 195 2/7 18%  C₁ 1.5 mg 7  57.8 ± 1.5127 2/7 9% C₂ 3 mg 7  90.2 ± 1.9 198 3/7 17%  D₁ 1.5 mg 7 128.5 ± 1.3282 4/7 5% D₂ 3 mg 7 115.2 ± 2.1 253 4/7 10%  E₁ 1.5 mg 7 130.5 ± 1.6287 4/7 4% E₂ 3 mg 7 121.2 ± 1.8 266 3/7 9% F₁ 1.5 mg 7 122.5 ± 1.7 2694/7 6% F₂ 3 mg 7 111.2 ± 1.9 244 3/7 11% 

The results showed that the prodrugs demonstrated strong antitumoractivity at 1.5 mg/kg dose and caused much less side effects (lessweight loss) when they were administered transdermally.

What is claimed is:
 1. A method for treating a condition in a biologicalsubject, comprising administrating to the biological subject a highpenetration compound having the following chemical structure:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein: T is selected Structure Na, Structure Nb, Structure Nc,Structure Ne, Structure Ng, Structure Nh, and Structure Nm:

wherein: R₁₁ and R₁₂ in Structure Na are same or different and eachindependently selected from H, substituted and unsubstituted alkyl,substituted and unsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl; R₁₁, R₁₂ and R₁₃ in Structure Nb, StructureNc, Structure Ng or Structure Nh are each independently substituted orunsubstituted alkylene; R₁₁ and R₁₃ in Structure Ne and Structure Nm areeach independently substituted or unsubstituted methine, and R₁₂ issubstituted or unsubstituted alkylene; each R₁₄ is selected from H,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted aryl, substituted and unsubstituted heteroaryl; L₁ isselected from a bond, O, S, -N(L₃)-, -NL₃)-CH₂—O, -NL₃)-CH₂-NL₃)-,CH₂—O—, -O-CHL₃)-O, and -S-CHL₃) L₂ is selected from a bond, O, S,-NL₃)-, -NL₃)-CH₂-O, -NL₃)-CH₂-NL₃)-, CH₂—O—, -O-CHL₃)-O, -S-CHL₃)-O-,-O-L₅-,-N-L₅-, -S-L₅- and -NL₃)-L₅-;L₄ is C═O or C═S; each L₃ is independently selected from H, substitutedand unsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted aryl, and substituted and unsubstituted heteroaryl; eachL₅ is independently selected from substituted and unsubstitutedalkylene, substituted and unsubstituted cycloalkylene, substituted andunsubstituted heterocycloalkylene, substituted and unsubstitutedarylene, and substituted and unsubstituted heteroarylene; F comprises amoiety of a nitrogen mustard compound, having a structure of StructureA:

is selected from Structure Y-a, Structure Y-b, and Structure Y-c:

is selected from substituted and unsubstituted aryl, Structure Ar-a,Structure Ar-b, Structure Ar-c, Structure Ar-d, Structure Ar-e,Structure Ar-f, Structure Ar-g, Structure Ar-h, and Structure Ar-i:

X₁ and X₂ are independently selected from Cl, Br, F, and OSO₂R₄, R₄ isselected from unsubstituted alkyl, substituted and unsubstitutedalkoxyl, substituted and unsubstituted perfluoroalkyl, substituted andunsubstituted alkyl halide, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl groups; X₃-X₇ are independentlyselected from NHCOR₄, OR₄, SR₄, NHR₄, OCOR₄, R₄, substituted andunsubstituted alkoxyl, substituted and unsubstituted alkylthio,substituted and unsubstituted alkylamino, substituted and unsubstitutedalkyl halide, H, F, Cl, Br, I, NO₂, CN, CF₃, NHCOCH₃, OCH₃, SCH₃, NH₂,NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅, OC₃H₇, CH₃, C₂H₅, and C₃H₇; z is aninteger; Y₁ is selected from CH₂, O, S, and NH; Y₂ and Y₃ are the sameor different and each is NHCOR₄, H, OH, NHCOCH₃, NHCOC₂H₅, Cl, F, Br, orI, or taken together is ═O; Y₄ is selected from R₄, CH₂, —(CH₂)n—, O, S,and NH; A is selected from amino acids, a-amino acids and p-amino acidsresidues; any CH₂ groups may be replaced with O, S, or NH; and when abond is not linked with any atom of an aryl or heteroaryl ring, the bondcan be put into any position of the ring.
 2. The method according toclaim 1, wherein the condition is selected from tumor and psoriasis. 3.The method according to claim 2, wherein the tumor is selected frombenign tumor, breast cancer, colon-rectum cancer, oral cancer, lung orother respiratory system cancers, skin cancers, uterus cancer,pancreatic cancer, prostate cancer, genital cancer, urinary organscancers, leukemia, and other blood and lymph tissues cancer.
 4. Themethod according to claim 1, wherein the high penetration compound isadministered to the biological subject through a route selected fromoral, enteral, buccal, nasal, topical, rectal, vaginal, aerosol,transmucosal, epidermal, transdermal, dermal, ophthalmic, pulmonary,subcutaneous, and parenteral administration.
 5. The method according toclaim 1, wherein the high penetration compound is selected from nitrogenmustards, nitrobenzyl mustards, phosphoramide mustard, isophosphoramidemustards and aldophosphamide.
 6. The method of claim 1, wherein the highpenetration compound is administered in a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier.
 7. The method of claim6, wherein the pharmaceutically acceptable carrier is polar.
 8. Themethod of claim 6, wherein the pharmaceutically acceptable carrier isselected from alcohol, acetone, ester, water, and aqueous solution, or acombination thereof.
 9. A method for treating psoriasis or a tumor in abiological subject, comprising administrating to the biological subjecta high penetration compound of Structure 1, or a stereoisomer or apharmaceutically acceptable salt thereof, wherein Structure 1 isselected from Structure 1a, Structure 1b, Structure 1c, Structure 1d,Structure 1e, Structure 1g, and Structure 1m, having the followingchemical structures:

wherein, in any of Structure 1a, Structure 1b, Structure 1c, Structure1d,

Structure 1 e, Structure 1g, and Structure 1m, is selected fromStructure Y-a, Structure Y-b, and Structure Y-c:

is selected from substituted and unsubstituted aryl, Structure Ar-a,Structure Ar-b, Structure Ar-c, Structure Ar-d, Structure Ar-e,Structure Ar-f, Structure Ar-g, Structure Ar-h and Structure Ar-i:

R₄ is selected from unsubstituted alkyl, substituted and unsubstitutedalkoxyl, substituted and unsubstituted perfluoroalkyl, substituted andunsubstituted alkyl halide, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl groups; X₃-X₄ are independentlyselected from NHCOR₅, OR₅, SRS, NHR₅, OCOR₅, R₅, substituted andunsubstituted alkoxyl, substituted and unsubstituted alkylthio,substituted and unsubstituted alkylamino, substituted and unsubstitutedalkyl halide, H, F, Cl, Br, I, NO₂, CN, CF₃, NHCOCH₃, OCH₃, SCH₃, NH₂,NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅, OC₃H₇, CH₃, C₂H₅, and C₃H₇; X₅-X₇ areindependently selected from NHCOR₄, OR₄, SR₄, NHR₄, OCOR₄, R₄,substituted and unsubstituted alkoxyl, substituted and unsubstitutedalkylthio, substituted and unsubstituted alkylamino, substituted andunsubstituted alkyl halide, H, F, Cl, Br, I, NO₂, CN, CF₃, NHCOCH₃,OCH₃, SCH₃, NH₂, NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅, OC₃H₇, CH₃, C₂H₅, andC₃H₇; z is an integer; Y₁ is selected from CH₂, O, S, and NH; Y₂ and Y₃are the same or different and each is NHCOR₄, H, OH, NHCOCH₃, NHCOC₂H₅,Cl, F, Br, or I, or taken together is ═O; Y₄ is selected from R₄, CH₂,—(CH₂)_(n)—, O, S, and NH; A is selected from amino acids, α-amino acidsand β-amino acids residues; any CH₂ groups may be replaced with O, S, orNH; and when a bond is not linked with any atom of an aryl or heteroarylring, the bond can be put into any position of the ring; R isindependently selected from substituted and unsubstituted 0-20 carbonatoms alkylene, substituted and unsubstituted 1-20 carbon atomsperfluoroalkylene, substituted and unsubstituted 2-20 carbon atomsalkenylene, substituted and unsubstituted 2-20 carbon atoms alkynylene,substituted and unsubstituted 6-20 carbon atoms arylene, and substitutedand unsubstituted 2-20 carbon atoms heteroarylene; R₁ and R₂ inStructure 1a are same or different and each independently selected fromH, substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted aryl, substituted and unsubstituted heteroaryl; R₁₁,and R₁₂ in Structure 1b and Structure 1c are each independentlysubstituted or unsubstituted alkylene; R₁₃ in Structure 1b are eachindependently substituted or unsubstituted alkylene; R₅ in Structure 1c,Structure 1e, Structure 1g, and Structure 1m is selected from H,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted aryl, substituted and unsubstituted heteroaryl; R₁₁and R₁₃ in Structure 1g and Structure 1m are each independentlysubstituted or unsubstituted methine, and R₁₂ is substituted orunsubstituted alkylene; X is selected from O, S, NRS, and NH; X₁ and X₂are independently selected from Cl, Br, F, and OSO₂R₅; and HA isselected from nothing, hydrochloride, hydrobromide, hydroiodide, nitricacid, sulfic acid, bisulfic acid, phosphoric acid, phosphorous acid,phosphonic acid, isonicotinic acid, acetic acid, lactic acid, salicylicacid, citric acid, tartaric acid, pantothenic acid, bitartaric acid,ascorbic acid, succinic acid, maleic acid, gentisinic acid, fumaricacid, gluconic acid, glucaronic acid, saccharic acid, formic acid,benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid,benzensulfonic acid, p-toluenesulfonic acid and pamoic acid.
 10. Themethod according to claim 9, wherein the tumor is selected from benigntumor, breast cancer, colon-rectum cancer, oral cancer, lung or otherrespiratory system cancers, skin cancers, uterus cancer, pancreaticcancer, prostate cancer, genital cancer, urinary organs cancers,leukemia, and other blood and lymph tissues cancer.
 11. The methodaccording to claim 9, wherein the high penetration compound isadministered to the biological subject through a route selected fromoral, enteral, buccal, nasal, topical, rectal, vaginal, aerosol,transmucosal, epidermal, transdermal, dermal, ophthalmic, pulmonary,subcutaneous, and parenteral administration.
 12. The method of claim 9,wherein the high penetration compound is administered in apharmaceutical composition comprising a pharmaceutically acceptablecarrier.
 13. The method of claim 12, wherein the pharmaceuticallyacceptable carrier is selected from alcohol, acetone, ester, water, andaqueous solution, or a combination thereof.
 14. A method for treatingpsoriasis or a tumor in a biological subject, comprising administratingto the biological subject a high penetration compound having thefollowing structure:

wherein: X₁ and X₂ are independently selected from Cl, Br, F, andOSO₂R₄; R₄ is selected from substituted and unsubstituted alkyl,substituted and unsubstituted alkoxyl, substituted and unsubstitutedperfluoroalkyl, substituted and unsubstituted alkyl halide, substitutedand unsubstituted aryl, and substituted and unsubstituted heteroarylgroups;

is selected from substituted and unsubstituted aryl, Structure Ar-a,Structure Ar-b, Structure Ar-c, Structure Ar-d, Structure Ar-e,Structure Ar-f, Structure Ar-g, Structure Ar-h and Structure Ar-i:

X₃-X₇ are independently selected from NHCOR₄, OR₄, SR₄, NHR₄, OCOR₄, R₄,substituted and unsubstituted alkoxyl, substituted and unsubstitutedalkylthio, substituted and unsubstituted alkylamino, substituted andunsubstituted alkyl halide, H, F, Cl, Br, I, NO₂, CN, CF₃, NHCOCH₃,OCH₃, SCH₃, NH₂, NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅, OC₃H₇, CH₃, C₂H₅, andC₃H₇; Y₁ is selected from CH₂, O, S, and NH; Y₂ and Y₃ are the same ordifferent and each is NHCOR₄, H, OH, NHCOCH₃, NHCOC₂H₅, Cl, F, Br, or I,or taken together is ═O; Y₄ is selected from R₄, CH₂, —(CH₂)n—, O, S,and NH; L₃ is an alkylene; R₁₁ and R₁₂ are independently selected fromH, substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted aryl, substituted and unsubstituted heteroaryl; and HAis selected from nothing, hydrochloride, hydrobromide, hydroiodide,nitric acid , sulfic acid, bisulfic acid, phosphoric acid, phosphorousacid, phosphonic acid, isonicotinic acid, acetic acid, lactic acid,salicylic acid, citric acid, tartaric acid, pantothenic acid, bitartaricacid, ascorbic acid, succinic acid, maleic acid, gentisinic acid,fumaric acid, gluconic acid, glucaronic acid, saccharic acid, formicacid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonicacid, benzensulfonic acid, p-toluenesulfonic acid and pamoic acid. 15.The method of claim 14, wherein the high penetration compound has thefollowing structure:

wherein: X₁ and X₂ are independently selected from Cl and OSO₂R₄; R₄ isselected from substituted and unsubstituted alkyl;

is selected from Structure Ar-a, Structure Ar-b, and Structure Ar-c:

X₃-X₆ are independently selected from NHCOR₄, OR₄, SR₄, NHR₄, OCOR₄, R₄,substituted and unsubstituted alkoxyl, substituted and unsubstitutedalkylthio, substituted and unsubstituted alkylamino, substituted andunsubstituted alkyl halide, H, F, Cl, Br, I, NO₂, CN, CF₃, NHCOCH₃,OCH₃, SCH₃, NH₂, NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅, OC₃H₇, CH₃, C₂H₅, andC₃H₇; Y₁ is selected from CH₂, O, S, and NH; Y₂ and Y₃ are the same ordifferent and each is NHCOR₄, H, OH, NHCOCH₃, NHCOC₂H₅, Cl, F, Br, or I,or taken together is ═O; Y₄ is selected from R₄, CH₂, -(CH₂)n-, O, S,and NH; L₃ is an alkylene; Ru and R₁₂ are independently selected from Hand substituted and unsubstituted alkyl; and HA is selected fromnothing, hydrochloride, hydrobromide, hydroiodide, nitric acid , sulficacid, bisulfic acid, phosphoric acid, phosphorous acid, phosphonic acid,isonicotinic acid, acetic acid, lactic acid, salicylic acid, citricacid, tartaric acid, pantothenic acid, bitartaric acid, ascorbic acid,succinic acid, maleic acid, gentisinic acid, fumaric acid, gluconicacid, glucaronic acid, saccharic acid, formic acid, benzoic acid,glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzensulfonicacid, p-toluenesulfonic acid and pamoic acid.
 16. The method of claim14, wherein the high penetration compound has the following structure:

wherein: X₁ and X₂ are independently selected from Cl and OSO₂R₄; R₄ isselected from substituted and unsubstituted alkyl;

is selected from Structure Ar-a, Structure Ar-b, and Structure Ar-c:

X₃-X₆ are independently selected from NHCOR₄, OR₄, SR₄, NHR₄, OCOR₄, R₄,substituted and unsubstituted alkoxyl, substituted and unsubstitutedalkylthio, substituted and unsubstituted alkylamino, substituted andunsubstituted alkyl halide, H, F, Cl, Br, I, NO₂, CN, CF₃, NHCOCH₃,OCH₃, SCH₃, NH₂, NHCH₃, OCOCH₃, OCOC₂H₅, OC₂H₅, OC₃H₇, CH₃, C₂H₅, andC₃H₇; Y₂ and Y₃ are the same or different and each is NHCOR₄, H, OH,NHCOCH₃, NHCOC₂H₅, Cl, F, Br, or I, or taken together is ═O; Y₄ isselected from R₄, CH₂, -(CH₂)n-, O, S, and NH; L₃ is an alkylene; R₁₁and R₁₂ are independently selected from H and substituted andunsubstituted alkyl; and HA is selected from nothing, hydrochloride,hydrobromide, hydroiodide, nitric acid , sulfic acid, bisulfic acid,phosphoric acid, phosphorous acid, phosphonic acid, isonicotinic acid,acetic acid, lactic acid, salicylic acid, citric acid, tartaric acid,pantothenic acid, bitartaric acid, ascorbic acid, succinic acid, maleicacid, gentisinic acid, fumaric acid, gluconic acid, glucaronic acid,saccharic acid, formic acid, benzoic acid, glutamic acid,methanesulfonic acid, ethanesulfonic acid, benzensulfonic acid,p-toluenesulfonic acid and pamoic acid.
 17. The method of claim 14,wherein the high penetration compound, or a pharmaceutically acceptablesalt thereof, is selected from:(4-[bis(2-chloroethyl)amino]-N-acetyl-L-phenylalanineN,N-diethylaminoethyl ester); (N,N-diethylaminoethyl4-[bis(2-chloroethyl)amino]benzenebutyrate); and (diethylaminoethyl4-[bis(2-methylsulfonylethyl)amino]benzenebutyrate).
 18. The methodaccording to claim 14, wherein the tumor is selected from benign tumor,breast cancer, colon-rectum cancer, oral cancer, lung or otherrespiratory system cancers, skin cancers, uterus cancer, pancreaticcancer, prostate cancer, genital cancer, urinary organs cancers,leukemia, and other blood and lymph tissues cancer.
 19. The methodaccording to claim 14, wherein the high penetration compound isadministered to the biological subject through a route selected fromoral, enteral, buccal, nasal, topical, rectal, vaginal, aerosol,transmucosal, epidermal, transdermal, dermal, ophthalmic, pulmonary,subcutaneous, and parenteral administration.
 20. The method of claim 14,wherein the high penetration compound is administered in apharmaceutical composition comprising a pharmaceutically acceptablecarrier selected from alcohol, acetone, ester, water, and aqueoussolution, or a combination thereof.